Concentrated enzymic substance and method of preparing same



Patented Dec. 8, 1925.

UNITED STATES PATENT OFFICE. i

SELMAN A. WAKSMAN, OF NEW. BRUNSWICK, NEW JERSEY, ASSIGE'OR TO JACQUESWOLF & CO., OF PASSAIG, NEW JERSEY. A CORPORATION OF NEW JERSEY.

CONCENTRATED ENZYMIO SUBSTANCE AND -METHOD OF PREPARING SAME.

No Drawing. Application filed August 11, 1922, Serial No. 581,236.Renewed February 28, 1925.

. stances and processes for producing the same and has for one of itsprincipal objects to provide a stable enzymic substance which is moreconcentrated than those heretofore pro-' duced, and "a method ofproducing the same- Enzymes are unorganized ferments, or chemicalsubstances of vegetable or animal origin that cause certain chemicaltransformationsv by their presence. Enzymes are produced by livingcells, either animal or vegetable, including various bacteria and molds,for example, by the propagation of mold fungi such as Aspergz'llus org2w or other mold fungi of the genera Aspergillus, Penicillium and Mucorupon various substances of plant or animal origin, including bran ofvarious grains. When brought into relation with certain organiccompounds, the action of the enzymes tends to decompose such compoundsinto simpler combinations without themselves undergoing theoreticallyany change, although practically the enzymes gradually disappear in thecourse of the reaction. Their precise mode of action is not definitelyunderstood but it is apparently catalytic. Enzymes act only in thepresence of", water and a small quantity serves to break up or change alarge mass of the substance or substances on which it acts. Enzymes havewell known properties such as diastatic and proteolytic. By virtue oftheir diastatic property, t ey may be employed to convert starch intosugar, this property being predominantly present in saliva which is anenzymic solution. By virtue of their proteolytlc property enzymes exerta digestive action upon meat and other proteins, this property beingpredominantly present in the enzymic digestive juices in the stomach andpancreas of human beings and animals.

The present invention has to do with enzymes of vegetable origin havingvarious ferent or separate mass of fungus-impreg nated culture medium isthen soaked or steeped in the liquid extract. The liquid is i thenevaporated, i. e., the mixture is dried, leaving the desired enzymicsubstance.

Preparation of the fungus-impregnated culture medium may be effected invarious ways. For example, it may be prepared in the manner disclosedin' the now expired United States Patents Nos. 525,820 or 525,823 inwhich the fungus-impregnated mass is given the name of taka-koji.

Another, and in many respects preferable method of producing thefungus-impregnated mass is substantiall as follows. bran, or otherequiva ent material; apple pomace, or another ventilating a ent; andwater are mixed, preferably in the following proportions: applepomace120 to 200 pounds; bran-enough to weigh with the pomaceabout 1000pounds; waterabout 50 allons. The mixture so formed is then sterilizedby steaming for about one hour and then cooled down to a temperature ofabout 30 C., about one hour being consumed in the cooling operation. Tothis mixture so prepared is added about 35 gallons of water containingthe seed spores. These seed spores are preferably those formed by moldfungi belonging to the genera Aspergillus, Penicillium or Mucor, such asthe WVheat species Aspergz'Z Zus oryzw (or Eurotz'wm.

Preparation of the liquid extract.

A dried, or undried, mass of fungus-impregnated culture medium islixiviated with water. This may be done by permitting ass in theliquid.- If desired, the strengtfi of water or other suitable liquid topercolate through the mass, or by steeping the this solution may beincreased by repeat dly treating a fresh quantity of the mass thererwith. The extract may then be filtered or or unstrained, constitutes thedesired exill ' last mentioned dry mixture out in a dry place.

koji. but in its enzymic tract. Water is added to the same batch ofculture medium and drained off to be employed as the liquid forlixiviating a succeeding'batoh of fresh culture medium.

The next step (is to soak or steep a fresh mass of culture described, inthe liquid extract, prepared as above described. The mixture is allowedto soak or steep for one pr two hours and then dried, i. e., the liquid1s evaporated off,,until the moisture content is reduced to 5 per centor less. The resultant novel substance is conveniently referred to asProtozyme concentrated. Further concentration of the desired productmay-be secured by stee ing or soaking the Protoz me concentrate pro:duced as described a ove, in a fresh quantity of liquid extract, againdrying-it, and if desirab e again repeating this process as many timesas desire Summarizing a preferred method of carrying out the process:two or more lots of Protozyme are prepared, each in the manner abovedescribed. One lot, is allowed to dry. The other lot or lots arelixiviated to form a liquid extract, as above described.

The lot of dry Protozyme isthen soaked in y the liquid extract and isthen dried. The product may then be steeped or soaked in afresh batch ofliquid extract, again dried, and this p ocess repeated as often asdesired, depending upon the degree of concentration de'siredfi Thedrying may be effected by an air current in a partial vacuum, or byexposure to the sun ona dry day, or by simply spreading the In any case,the drying is continued until the moisture content is reduced to 5 percent or less. The resultant dry product is Protozyme concentrated.

This resultant product not only ossesses the stabilityAof dry Protozymeor takaproperties (diastatic and proteolytic and others) it is more thantwice as concentrated as either of its constituents-Protozyme and liquidextract of Protozyme. v

medium, prepared as above It has been proposed by various workers in theenzymic art (see pages 769 and 770 of Allegemeine Mikrobiologie, Kruse1910, published at Leipzig, Germany) to preserve enzymic solutions byemploying antiseptics of various kinds. In my process I employ noantiseptic whatsoever, yet the resultant product does not deterioratebecause it is in dry form.

The novel product resulting from the above described novel process maybe described as a dry enzymic substancecomprising a fungus-impregnatedculture medium and the concentrate (i. e., a product of a process ofconcentration by drying or evaporation) of a liquid extract of afungus-impregnated culture medium. This product is powerful in itsenzymic properties, though of small bulk. Being dry, i. e.,of reducedmoisture content, it will keep and preserve its enzymic properties,practically indefinitely.

The novel product maybe employed to great advantage in the clarificationof fruit juices, as a medicinal preparation against dyspepsia and invarious other fields, arts and industries.

What I claim is:

1. The process of preparingan enzymic substance gas-impregnated culturemedium, soaking in the liquid extract a fresh mass of fungusimpregnatedculture medim, and drying the mixture.

2. The process of preparing an enzymi'c substance which comprisespreparing two lots offungus-impregnated culture medium, drying out onelot, lixiviating the other lot to form an aqueous extract, soaking thedried lot in the extract formed from the other lot, and drying themixture.

3. The process of preparing an enzymic substance which comprisespropagating a fungus on a culture medium, drying one lot of such medium,lixiviating another lot of such medium with a liquid to which a thirdlot of culturemedi'um has been subjected, soaking the dried lot ofculture medium in the extract, and reducing the moisture con tent of themixture until not more than 5 per cent moisture remains. 1 4. As a newarticle of manufacture, a dry enzymic substance comprising afungus-impregnated culture medium and the concen trate of a liquidextract of a fungus-impregnated culture medium. a

5. The process of preparing an enzymic substance Which compriseslixiviating a funthe mixture.

impregnated culture medium, and drying 13o 6. As a new article ofmanufacture, a. ry pregnated culture medium and the concenenzymicsubstance comprising a fungus-' trates of liquid extracts of afungus-impregpre ated culture med1um and the concennated culture medium.trate'of a liquid extract of difl'erent lots of In testimony whereof Ihereto afiix my 5 fungus-impregnated culture medium. signature.

7. As a new article of manufacture, a dry enzymic substance comprising afungus-im- SELMAN A. WAKSMAN.

